LOCUS       62056_21795        9289 bp DNA     circular SYN 26-FEB-2020
DEFINITION  Cloning vector pUASGal80-PUbEGFP-attB, complete sequence.
ACCESSION   MN830806
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9289)
  AUTHORS   Webster SH, Vella MR, Scott MJ.
  TITLE     Development and testing of a novel Killer-Rescue self-limiting gene 
            drive system in Drosophila melanogaster
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 9289)
  AUTHORS   Webster SH, Vella MR, Scott MJ.
  TITLE     Direct Submission
  JOURNAL   Submitted (13-DEC-2019) Entomology and Plant Pathology, North 
            Carolina State University, 112 Derieux Place, 1546 Thomas Hall, 
            Raleigh, NC 27607, USA
REFERENCE   3  (bases 1 to 9289)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (13-DEC-2019) Entomology and Plant Pathology, North Carolina State 
            University, 112 Derieux Place, 1546 Thomas Hall, Raleigh, NC 27607, 
            USA"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..9289
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        243..2051
                     /label=Polyubiquitin
                     /note="Promoter of polyubiqutin gene of Drosophila
                     melanogaster"
     CDS             2076..2096
                     /label=SV40 NLS
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
     CDS             2280..2996
                     /label=EGFP
                     /note="enhanced GFP"
     polyA_signal    3122..3243
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        complement(3309..3327)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        3358..3376
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(3415..3433)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3440..3456)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      complement(3597..4052)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        4079..4183
                     /label=AmpR promoter
     CDS             4184..5041
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      5215..5803
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    6091..6112
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        6127..6157
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6165..6181
                     /label=lac operator
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     6189..6205
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6226..6244
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     protein_bind    complement(6463..6532)
                     /label=attB
                     /note="attB site for the phi-C31 integrase (Groth et al.,
                     2000)"
     protein_bind    6669..6763
                     /label=5X UAS
                     /note="five tandem copies of the 'ScaI site' 17-mer 
                     CGGAGTACTGTCCTCCG, an upstream activating sequence (UAS) 
                     that efficiently binds yeast Gal4 (Webster et al., 1988; 
                     Pfeiffer et al., 2010)"
     promoter        6922..7160
                     /label=hsp70 promoter
                     /note="Drosophila melanogaster hsp70Bb promoter"
     intron          7175..7241
                     /label=Mhc intron
                     /note="modified Drosophila myosin heavy chain intron 16 
                     (Pfeiffer et al., 2010)"
     regulatory      7264..7284
                     /label=syn21 translation enhancer
                     /note="syn21 translation enhancer"
                     /regulatory_class="other"
     CDS             7285..8589
                     /label=GAL80
                     /note="transcriptional repressor that inhibits GAL4"
     regulatory      8598..9264
                     /label=p10 pA
                     /note="p10 pA"
                     /regulatory_class="polyA_signal_sequence"
ORIGIN
        1 atcgatcgcg cgcagatctg tcatgatgat cattgcaatt ctgcagtcga cggtacccga
       61 tcttgtcgcc ggaacgcagc gacagagatt ccaatgtgtc cgtatctttc aggcttttgc
      121 ccttcagttc cagacgaagc gactggcgat tcgcgtgtgg ggtctgcttc agggtcttgt
      181 gaattagggc gcgcagatcg ccgatgggcg tggcgccgga gggcaccttc accttgccgt
      241 acggcttgct gttcttcgcg ttcaaaatct ccagctccat tttgctttcg gtgcgcttgc
      301 aatcagtact gtccaaaatc gaaaatcgcc gaaccgtagt gtgaccgtgc ggggctctgc
      361 gaaaataaac ttttttaggt atatggccac acacggggaa agcacagtgg attatatgtt
      421 ttaatattat aatatgcagg ttttcattac ttatccagat gtaagcccac ttaaagcgat
      481 ttaacaatta tttgccgaaa gagtaaaaac aaatttcact taaaaatgga ttaagaaaag
      541 cttgtgtaag attatgcgca gcgttgccag atagctccat ttaaaacact tcaaaaacaa
      601 taagttttga aaatatatac ataaatagca gtcgttgccg caacgctcaa cacatcacac
      661 ttttaaaaca ccctttacct acacagaatt actttttaaa tttccagtca agctgcgagt
      721 ttcaaaatta tagccggtag agaagacagt gctatttcaa aagcaaacta aataaacacc
      781 aatcctaaca agccttggac ttttgtaagt ttagatcaaa ggtggcattg cattcaatgt
      841 catggtaaga agtaggtcgt ctaggtagaa atcctcattc agccggtcaa gtcagtacga
      901 gaaaggtctc aatttgaaat tgtcttaaaa atattttatt gttttgtact gtggtgagtt
      961 taaacgaaaa acacaaaaaa aaagtgatac acagaaatca taaaaaattt taatacaagg
     1021 tattcgtacg tatcaaaaac atttcggcac aatttttttt ctctgtacta aagtgttacg
     1081 aacactacgg tattttttag tgattttcaa cggacaccga aggtatataa acagcgttcg
     1141 cgaacggtcg ccttcaaaac caattgacat ttgcagcagc aagtacaagc agaaagtaaa
     1201 gcgcaatcag cgaaaaattt atacttaatt gttggtgatt aaagtacaat taaaagaaca
     1261 ttctcgaaag tcacaagaaa cgtaagtttt taactcgctg ttaccaatta gtaataagag
     1321 caacaagacg ttgagtaatt tcaagaaaaa ctgcatttca aggtctttgt tcggccattt
     1381 tttttttatt caacgctcta cgtaattaca aaataagaaa ttggcagcca cgcatcttgt
     1441 tttcccaatc aattggcatc aaaacgcaaa caaatctata aataaaactt gcgtgttgat
     1501 tttcgccaag atttattggc aaattgtgaa attcgcagtg acgcatttga aaattcgaga
     1561 aatcacgaac gcactcgagc atttgtgtgc atgttattag ttagttagtt ctttgcttaa
     1621 ttgaagtatt ttaccaacga aatccactta tttttagctg aaatagagta ggttgcttga
     1681 aacgaaagcc acgtctggaa aatttcttat tgcttagtag ttgtgacgtc accatataca
     1741 cacaaaataa tgtgtatgca tgcgtttcag ctgtgtatat atacatgcac acactcgcat
     1801 tatgaaaacg atgacgagca acggaacagg tttctcaact acctttgttc ctgtttcttc
     1861 gctttccttt gttccaatat tcgtagaggg ttaatagggg tttctcaaca aagttggcgt
     1921 cgataaataa gtttcccatt tttattcccc agccaggaag ttagtttcaa tagttttgta
     1981 atttcaacga aactcatttg atttcgtact aattttccac atctctattt tgacccgcag
     2041 aataatccaa aatgcagatc ggggatccca ccccacccaa gaagaagcgc aaggtggagg
     2101 acgatcccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc
     2161 gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc
     2221 gcccttccca acagttgcgg tcgactctag aggatccccg ggatccaccg gtcgccacca
     2281 tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg
     2341 gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg
     2401 gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc
     2461 tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc
     2521 agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct
     2581 tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg
     2641 tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca
     2701 agctggagta caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg
     2761 gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg
     2821 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact
     2881 acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc
     2941 tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtaaa
     3001 gcggccgcga ctctagatca taatcagcca taccacattt gtagaggttt tacttgcttt
     3061 aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt
     3121 taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     3181 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     3241 ttaaagctta tcgatacgcg tacggcacta gagcggccgc caccgcggtg gagctccagc
     3301 ttttgttccc tttagtgagg gttaattaga tcggccggcc ttggcgcgcc tagatcttaa
     3361 tacgactcac tatagggcga attgggtacc gggcccggta cgtacccaat tcgccctata
     3421 gtgagtcgta ttacaattca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg
     3481 gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg
     3541 aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatggaaat
     3601 tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
     3661 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg
     3721 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
     3781 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc
     3841 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg
     3901 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
     3961 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc
     4021 cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg
     4081 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     4141 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     4201 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     4261 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     4321 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     4381 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     4441 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     4501 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     4561 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     4621 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     4681 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     4741 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     4801 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     4861 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     4921 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     4981 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     5041 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     5101 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     5161 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     5221 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     5281 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     5341 agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa
     5401 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     5461 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     5521 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     5581 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     5641 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     5701 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     5761 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     5821 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     5881 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     5941 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa
     6001 accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga
     6061 ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc
     6121 ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca
     6181 atttcacaca ggaaacagct atgaccatga ttacgccaag ctcgaaatta accctcacta
     6241 aagggaacaa aagctggcta gaactagtgt cgacatgccc gccgtgaccg tcgagaaccc
     6301 gctgacgctg ccccgcgtat ccgcacccgc cgacgccgtc gcacgtcccg tgctcaccgt
     6361 gaccaccgcg cccagcggtt tcgagggcga gggcttcccg gtgcgccgcg cgttcgccgg
     6421 gatcaactac cgccacctcg acccgttcat catgatggac cagatgggtg aggtggagta
     6481 cgcgcccggg gagcccaagg gcacgccctg gcacccgcac cgcggcttcg agaccgtgac
     6541 ctacatcgtc gacactagtg gatccagcgg ccgcacctgc aggccggccg ttaacacgcg
     6601 tccgcggtct agactcgagg attccagatc tggtaccgct agtgcgtcaa gcttgcatgc
     6661 ctgcaggtcg gagtactgtc ctccgagcgg agtactgtcc tccgagcgga gtactgtcct
     6721 ccgagcggag tactgtcctc cgagcggagt actgtcctcc gagcggagac tctagcccta
     6781 gggcatgcct gcaggtcgga gtactgtcct ccgagcggag tactgtcctc cgagcggagt
     6841 actgtcctcc gagcggagta ctgtcctccg agcggagtac tgtcctccga gcggagactc
     6901 tagcgctagc gacgtcgagc gccggagtat aaatagaggc gcttcgtcta cggagcgaca
     6961 attcaattca aacaagcaaa gtgaacacgt cgctaagcga aagctaagca aataaacaag
     7021 cgcagctgaa caagctaaac aatctgcagt aaagtgcaag ttaaagtgaa tcaattaaaa
     7081 gtaaccagca accaagtaaa tcaactgcaa ctactgaaat ctgccaagaa gtaattattg
     7141 aatacaagaa gagaactctg aatagatcta aaaggtaggt tcaaccactg atgcctaggc
     7201 acaccgaaac gactaaccct aattcttatc ctttacttca ggcggccgcg gctcgagggt
     7261 accaacttaa aaaaaaaaat caaaatggac tacaacaaga gatcttcggt ctcaaccgtg
     7321 cctaatgcag ctcccataag agtcggattc gtcggtctca acgcagccaa aggatgggca
     7381 atcaagacac attaccccgc catactgcaa ctatcgtcac aatttcaaat cactgcctta
     7441 tacagtccaa aaattgagac ttctattgcc accattcagc gtctaaaatt gagtaatgcc
     7501 actgcttttc ccactttaga gtcatttgca tcatcttcca ctatagatat gatagtgata
     7561 gctatccaag tggccagcca ttatgaagtt gttatgcctc tcttggaatt ctccaaaaat
     7621 aatccgaacc tcaagtatct tttcgtagaa tgggcccttg catgttcact agatcaagcc
     7681 gaatccattt ataaggctgc tgctgaacgt ggggttcaaa ccatcatctc tttacaaggt
     7741 cgtaaatcac catatatttt gagagcaaaa gaattaatat ctcaaggcta tatcggcgac
     7801 attaattcga tcgagattgc tggaaatggc ggttggtacg gctacgaaag gcctgttaaa
     7861 tcaccaaaat acatctatga aatcgggaac ggtgtagatc tggtaaccac aacatttggt
     7921 cacacaatcg atattttaca atacatgaca agttcgtact tttccaggat aaatgcaatg
     7981 gttttcaata atattccaga gcaagagctg atagatgagc gtggtaaccg attgggccag
     8041 cgagtcccaa agacagtacc ggatcatctt ttattccaag gcacattgtt aaatggcaat
     8101 gttccagtgt catgcagttt caaaggtggc aaacctacca aaaaatttac caaaaatttg
     8161 gtcattgaca ttcacggtac caagggagat ttgaaacttg aaggcgatgc cggcttcgca
     8221 gaaatttcaa atctggtcct ttactacagt ggaactagag caaacgactt cccgctagcc
     8281 aatggacaac aagctccttt agacccgggg tatgatgcag gtaaagaaat catggaagta
     8341 tatcatttac gaaattataa tgccattgtg ggtaatattc atcgactgta tcaatctatc
     8401 tctgacttcc acttcaatac aaagaaaatt cctgaattac cctcacaatt tgtaatgcaa
     8461 ggtttcgatt tcgaaggctt tcccaccttg atggatgctc tgatattaca caggttaatc
     8521 gagagcgttt ataaaagtaa catgatgggc tccacattaa acgttagcaa tatctcgcat
     8581 tatagtttat aatctagaat gaatcgtttt taaaataaca aatcaattgt tttataatat
     8641 tcgtacgatt ctttgattat gtaataaaat gtgatcatta ggaagattac gaaaaatata
     8701 aaaaatatga gttctgtgtg tataacaaat gctgtaaacg ccacaattgt gtttgttgca
     8761 aataaaccca tgattatttg attaaaattg ttgttttctt tgttcataga caatagtgtg
     8821 ttttgcctaa acgtgtactg cataaactcc atgcgagtgt atagcgagct agtggctaac
     8881 gcttgcccca ccaaagtaga ttcgtcaaaa tcctcaattt catcaccctc ctccaagttt
     8941 aacatttggc cgtcggaatt aacttctaaa gatgccacat aatctaataa atgaaataga
     9001 gattcaaacg tggcgtcatc gtccgtttcg accatttccg aaaagaactc gggcataaac
     9061 tctatgattt ctctggacgt ggtgttgtcg aaactctcaa agtacgcagt caggaacgtg
     9121 cgcgacatgt cgtcgggaaa ctcgcgcgga aacatgttgt tgtaaccgaa cgggtcccat
     9181 agcgccaaaa ccaaatctgc cagcgtcaat agaatgagca cgatgccgac aatggagctg
     9241 gcttggatag cgattcgagt taacggccgg ccgtttaaac gaattcgat
//
